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transposase pbase  (Amaxa)

 
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    Amaxa transposase pbase
    Transposase Pbase, supplied by Amaxa, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+pbase/pbase+transposase/pmc13138334-126-19-27
    Average 86 stars, based on 1 article reviews
    transposase pbase - by Bioz Stars, 2026-10
    86/100 stars

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    Related Articles

    Transgenic Assay:

    Article Title: The conserved N-terminal SANT1-binding domain (SBD) of EZH2 regulates PRC2 activity.
    Article Snippet: .. To generate transgenic mouse ES cell lines, ∼5×106 cells were electroporated with PiggyBac expression vectors plus transposase (pBase) in a 3:1 ratio using the Amaxa ESC nucleofector kit (Lonza VPH-1001, program A-023). ..

    Article Title: The conserved N-terminal SANT1-binding domain (SBD) of EZH2 regulates PRC2 activity
    Article Snippet: To generate knockout lines, mouse ES cells were transfected with sgRNA-containing PX458 using Xfect mouse ES cell transfection reagent (Takara) and incubated for 48 h. Single GFP + cells were then sorted into 96 well plates. .. To generate transgenic mouse ES cell lines, ∼5 × 10 6 cells were electroporated with PiggyBac expression vectors plus transposase (pBase) in a 3:1 ratio using the Amaxa ESC nucleofector kit (Lonza VPH-1001, program A-023). .. To generate transgenic mouse ES cell lines, ∼5 × 10 6 cells were electroporated with PiggyBac expression vectors plus transposase (pBase) in a 3:1 ratio using the Amaxa ESC nucleofector kit (Lonza VPH-1001, program A-023).

    Expressing:

    Article Title: The conserved N-terminal SANT1-binding domain (SBD) of EZH2 regulates PRC2 activity.
    Article Snippet: .. To generate transgenic mouse ES cell lines, ∼5×106 cells were electroporated with PiggyBac expression vectors plus transposase (pBase) in a 3:1 ratio using the Amaxa ESC nucleofector kit (Lonza VPH-1001, program A-023). ..

    Article Title: The conserved N-terminal SANT1-binding domain (SBD) of EZH2 regulates PRC2 activity
    Article Snippet: To generate knockout lines, mouse ES cells were transfected with sgRNA-containing PX458 using Xfect mouse ES cell transfection reagent (Takara) and incubated for 48 h. Single GFP + cells were then sorted into 96 well plates. .. To generate transgenic mouse ES cell lines, ∼5 × 10 6 cells were electroporated with PiggyBac expression vectors plus transposase (pBase) in a 3:1 ratio using the Amaxa ESC nucleofector kit (Lonza VPH-1001, program A-023). .. To generate transgenic mouse ES cell lines, ∼5 × 10 6 cells were electroporated with PiggyBac expression vectors plus transposase (pBase) in a 3:1 ratio using the Amaxa ESC nucleofector kit (Lonza VPH-1001, program A-023).



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    Evaluation of <t>PiggyBac</t> transposon integration. A, Time course for evaluation of PiggyBac transposon (PB transposon) integration. The day when the donor vector carrying EGFP flanked by PiggyBac inverted terminal repeats (ITRs) and the effector vector carrying mammalian codon-optimised PiggyBac <t>transposase</t> (mPBase) were transfected at several concentrations was defined as Day 0. Fluorescence-activated cell sorting (FACS) was conducted every 2 days after transfection to confirm the ration of EGFP-positive mES cells. Passages were conducted using the remaining murine embryonic stem (mES) cells after FACS. B, Ratio of EGFP-positive mES cells and days after transfection. EGFP-positive mES cells were calculated among a total of 200 mES cells passing quality filters by FACS. Blank symbols in the left figure indicate mES cells transfected with 200 ng of donor vector, and filled symbols in the right figure indicate those transfected with 2,000 ng of donor vector. Gray circles, black squares, blue diamonds, and orange triangles indicate mES cells transfected with 0, 350, 3500, and 17500 ng of effector vector, respectively. We stopped evaluating mES cells transfected with a very high concentration (17500 ng) of effector vector on Day 4. C, Histogram of EGFP-positive mES cells on Day 10. The upper table shows the conditions of transfection. Lower histograms show all EGFP-positive mES cells in a total of 200 mES cells passing quality filters. The x-axis shows EGFP signal intensity, and the y-axis shows counts of EGFP-positive mES cells.
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    Evaluation of PiggyBac transposon integration. A, Time course for evaluation of PiggyBac transposon (PB transposon) integration. The day when the donor vector carrying EGFP flanked by PiggyBac inverted terminal repeats (ITRs) and the effector vector carrying mammalian codon-optimised PiggyBac transposase (mPBase) were transfected at several concentrations was defined as Day 0. Fluorescence-activated cell sorting (FACS) was conducted every 2 days after transfection to confirm the ration of EGFP-positive mES cells. Passages were conducted using the remaining murine embryonic stem (mES) cells after FACS. B, Ratio of EGFP-positive mES cells and days after transfection. EGFP-positive mES cells were calculated among a total of 200 mES cells passing quality filters by FACS. Blank symbols in the left figure indicate mES cells transfected with 200 ng of donor vector, and filled symbols in the right figure indicate those transfected with 2,000 ng of donor vector. Gray circles, black squares, blue diamonds, and orange triangles indicate mES cells transfected with 0, 350, 3500, and 17500 ng of effector vector, respectively. We stopped evaluating mES cells transfected with a very high concentration (17500 ng) of effector vector on Day 4. C, Histogram of EGFP-positive mES cells on Day 10. The upper table shows the conditions of transfection. Lower histograms show all EGFP-positive mES cells in a total of 200 mES cells passing quality filters. The x-axis shows EGFP signal intensity, and the y-axis shows counts of EGFP-positive mES cells.

    Journal: bioRxiv

    Article Title: Regional random mutagenesis driven by multiple sgRNAs and diverse on-target genome editing events to identify functionally important elements in non-coding regions

    doi: 10.1101/2023.09.12.557308

    Figure Lengend Snippet: Evaluation of PiggyBac transposon integration. A, Time course for evaluation of PiggyBac transposon (PB transposon) integration. The day when the donor vector carrying EGFP flanked by PiggyBac inverted terminal repeats (ITRs) and the effector vector carrying mammalian codon-optimised PiggyBac transposase (mPBase) were transfected at several concentrations was defined as Day 0. Fluorescence-activated cell sorting (FACS) was conducted every 2 days after transfection to confirm the ration of EGFP-positive mES cells. Passages were conducted using the remaining murine embryonic stem (mES) cells after FACS. B, Ratio of EGFP-positive mES cells and days after transfection. EGFP-positive mES cells were calculated among a total of 200 mES cells passing quality filters by FACS. Blank symbols in the left figure indicate mES cells transfected with 200 ng of donor vector, and filled symbols in the right figure indicate those transfected with 2,000 ng of donor vector. Gray circles, black squares, blue diamonds, and orange triangles indicate mES cells transfected with 0, 350, 3500, and 17500 ng of effector vector, respectively. We stopped evaluating mES cells transfected with a very high concentration (17500 ng) of effector vector on Day 4. C, Histogram of EGFP-positive mES cells on Day 10. The upper table shows the conditions of transfection. Lower histograms show all EGFP-positive mES cells in a total of 200 mES cells passing quality filters. The x-axis shows EGFP signal intensity, and the y-axis shows counts of EGFP-positive mES cells.

    Article Snippet: The PiggyBac transposase (PBase) effector vector (pCX-CAG>mPBase-PGK>HSV-TK) was constructed as follows: a custom fragment containing a Kozak sequence attached to the coding sequence (CDS) of mammalian codon-optimised PBase (mPBase) (GenBanK: EF587698.1 ) was synthesised by Integrated DNA Technologies (Coralville, IA).

    Techniques: Plasmid Preparation, Transfection, Fluorescence, FACS, Concentration Assay